Para-xylene films and therapeutic uses thereof

ABSTRACT

The present invention provides single sheet and compound para-xylene films for therapeutic uses. For example, the present invention provides single sheet para-xylene films useful as tissue separators and/or adhesion barriers in a subject, where the top and/or bottom surfaces of such films have a water contact angle between 75 and 95 degrees (e.g., to prevent adhesion formation). The present invention also provides compound films composed of at least two para-xylene polymer films with a therapeutic molecule layer in between. Such compound films, when used in vivo (e.g., as a tissue separator and to treat inflammation or atrial fibrillation) allow either therapeutic molecule elution through one of the para-xylene layers, or therapeutic molecule release when the compound film is pierced, such as when it is sutured in place.

The present application is a continuation of U.S. patent applicationSer. No. 14/283,819, filed May 21, 2014, allowed, which claims priorityto U.S. Patent Application Ser. No. 61/825,775 filed May 21, 2013, eachof which is herein incorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention provides for a tunable non-adherent and/oradherent single sheet, film or coating compounds comprising orconsisting of para-xylene derived polymers for therapeutic uses.

BACKGROUND

Post-surgical adhesions are generally fibrous bands that form betweentissues and organs as a result of surgery. They may be thought of asinternal scar tissue that connect tissues not normally connected, orconnect permanent or resorbable implants to adjacent tissue and/or organsurfaces in a manner which is detrimental to device and/or organfunction, and/or internal tissue, organ, or organ system spatialarrangement. Incorporation or integration of permanent implants to aspecific tissue surface or organ may be desired however.

Adhesions form as a natural part of the body's healing process aftersurgery in the same way that a scar forms. The term adhesion isgenerally applied when the scar extends from within one tissue across toanother, usually across a virtual space such as the peritoneal orthoracic cavity. As part of the process, the body deposits fibrin ontoinjured tissues. The fibrin acts like a glue to seal the injury andbuilds the fledgling adhesion, said at this point to be “fibrinous.” Inbody cavities such as the peritoneal, pericardial and synovial cavities,a family of fibrinolytic enzymes may act to limit the extent of theinitial fibrinous adhesion, and may even dissolve it. In many caseshowever the production or activity of these enzymes are compromisedbecause of injury, and the fibrinous adhesion persists. If this isallowed to happen, tissue repair cells such as macrophages, fibroblastsand blood vessel cells, penetrate into the fibrinous adhesion, and laydown collagen and other matrix substances to form a permanent fibrousadhesion. While some adhesions do not cause problems, others can preventmuscle and other tissues and organs from moving freely, sometimescausing organs to become twisted or pulled from their normal positions.

Adhesions can form in the thoracic cavity, such as after cardiac surgeryor related procedures. After cardiac surgery, inflammation from surgicaltrauma triggers adverse events. These include fluid retention, weightgain, pleural or pericardial effusions, pulmonary congestion,pericarditis, “postcardiotomy syndrome,” and new-onset atrialfibrillation (with the potential for hemodynamic compromise andstroke).¹⁻⁴ In addition, the length of hospitalization as well as theutilization of medical resources may be increased (such as the need forpleural effusion drainage and therapy for atrial fibrillation), andpatients are at increased risk for readmission. These complications andtreatments may add significantly to the cost of care. Medical therapiesto prevent these complications have had only modest success.⁵⁻⁸Therapies targeted to inhibit adhesions and the inflammatory responseare needed.

SUMMARY OF THE INVENTION

In certain embodiments, the present invention provides for a transparenttunable non-adherent and/or adherent single sheet, film or coatingcompounds comprising or consisting of non-degradable/non-resorbablepara-xylene derived polymers for therapeutic uses (e.g., to modulatecell and/or tissue response through interaction/adherence and/orintegration to said polymer surface and act as a drug deliveryplatform).

In certain embodiments, the present invention provides for a tunablenon-adherent and/or adherent single sheet, film or coating compoundcomprising, consisting essentially of, or consisting of para-xylenederived polymers (e.g., for therapeutic uses). For example, the presentinvention provides single sheet/film and/or coating of para-xylenepolymer(s) useful as tissue separators, tissue adherent platforms and/oradhesion barriers in a subject, where the top and/or bottom surfaces ofsuch films and/or coatings have a tunable chemical structure containingpredominately carbon related groups, alipathic compounds, including, butnot limited to, aromatic hydrocarbons with a benzene derived backbonevarying oxygen or nitrogen content to either prevent or promote cell ortissue adherence to said film or coating. Tuning of surface chemicalcontent lend to a variety of applications where certain anti-adhesiveand/or adhesive characteristics can be employed either as a film or as acoating, with thickness ranging from about 1 μm to 75 μm, or from 0.1 umto 0.2 mm (e.g., 0. 1 um . . . 1 um . . . 50 um . . . 0.1 mm . . . 0.2mm) to prevent common post-surgical complications relating to, forexample, scarring, surgical adhesion, bowel obstruction, and any numberof hernia or herniation related disorders.

In particular embodiments, anti-adherent surfaces are characterized byCarbon (C) and Oxygen (O) content of para-xylene surfaces with C to Oratios ranging from about 2:1 to 18:1, and/or water contact angles of75-95 and/or surface roughness values of 1.0-10.0 nm.

In other embodiments, adherent para-xylene surfaces are characterized bya decrease in water contact angles ranging from about 1-75, roughnessvalues of 1.0-10.0 nm, and resultant variation in C to O ratios lessthan 2:1 (e.g., through oxidative etching of the surface resulting inthe formation of predominantly carbonyl groups, to a lesser extentcarboxyl groups, and/or incorporation of nitrogen containing groupswhich will elicit a strong adherence and incorporation of cellular andtissue surfaces to said para-xylene sheet/film and/or surface/coating).The present invention also provides compound films composed of at leasttwo para-xylene polymer films with a therapeutic molecule layer and/orwireless device and/or sensor in between. Such compound films, when usedin vivo (e.g., as a tissue separator and to treat inflammation or atrialfibrillation) allow either therapeutic molecule elution through one ofthe para-xylene layers, and/or therapeutic molecule release when thecompound film is pierced, such as when it is sutured in place.Integrated or incorporated wireless device and/or sensor would be totrack certain patient environmental or physical conditions, wherein saidsensor would then relay patient related information wirelessly when areceiver or recorder is placed on or near the implant and/or patient.

In some embodiments, the present invention provides articles ofmanufacture comprising, consisting of, or consisting essentially of: apolymer film and/or coating (e.g., which is transparent) with a topsurface and a bottom surface, wherein the polymer film comprises,consists of, or consists essentially of, a para-xylene polymer, whereinthe polymer film has a thickness between 1 μm and 5 mm (e.g., 1. . . 5 .. . 15 . . . 25 . . . 38 . . . 55 . . . 75 μm . . . 1 mm . . . 3 mm 5mm), wherein the top surface has a water contact angle between 75 and 95degrees (e.g., 75 . . . 79 . . . 81 . . . 84 . . . 88 . . . 91 . . . or95 degrees), and/or a surface roughness of 1.0 nm and 10.0 nm (e.g. 1.0. . . 3.0 . . . 5.0 . . . or 10.0 nm), and/or a Carbon (C) to Oxygen (O)ratio of 2:1 and 18:1 (e.g., 2:1 . . . 6:1 . . . 10:1 . . . 14:1 . . .18:1) to create anti-adherent surfaces, wherein the polymer film and/orcoating is sized to serve as a tissue separator and/or adhesion barrierin a subject. For adherent surfaces, a water contact angle between 1-75degrees may be employed (e.g., 1 . . . 15 . . . 30 . . . 45 . . . 60 . .. 75) and C to O ratios less than 2:1 which may include the presence ofcarbonyl, carboxyl, and nitrogen containing groups to create adherentsurfaces wherein the polymer film and/or coating is sized to serve as atissue separator, tissue adherent platform and/or adhesion barrier in asubject. In certain embodiments, the polymer film has an opticaltransmission of 60-99.9% in the high visible (400-1000 nm) and nearinfra-red (3200-6000 cm) range.

In certain embodiments, the present invention provides methods forseparating areas in an internal region of a subject comprising:inserting an article of manufacture into an internal region of a subjectbetween first and second areas, wherein the article of manufactureconsists of, or consists essentially of, a polymer film and/or coatingwith a top surface and a bottom surface, wherein the polymer filmcomprises, consists of, or consists essentially of, a para-xylenepolymers, wherein the polymer film and/or coating has a thicknessbetween 1 μm and 5 mm (e.g., 1 . . . 5 . . . 15 . . . 25 . . . 38 . . .55 . . . 75 μm . . . 1 mm 5 mm), and wherein the top surface has a watercontact angle between 75 and 95 degrees (e.g., 75 . . . 79 . . . 81 . .. 84 . . . 88 . . . 91 . . . or 95 degrees). In certain embodiments, thepolymer film is contained and/or inserted into or within a Teflonflap/envelope or other component acting to hold said polymer film inplace, this combination of materials, in certain embodiments, is furthercontained or inserted into a final secondary packaging, which is sealedand then sterilized.

In certain embodiments, provided herein are the creation of individualor multiple adherence points, or a completely adherent side, to saidpara-xylene films and/or coatings. The function of these points would beto adhere cellular tissue and/or organ surfaces to a point, portion, orside of the para-xylene film. The number, size, and extent of theseadherence points would be dependent on the application. In particularembodiments, these areas would also be limited in scope or size suchthat they are sufficient to facilitate film adherence or attachment in apermanent manner to one portion or side of a tissue/organ surface orbody cavity lining.

In particular embodiments, manufacture of said adherence points, areas,or side would be through some method of oxidation, oxygen plasmaetching, ambient air plasma etching, and/or exposure, or UV irradiation,or any combination thereof which would impart primarily carbonylfunctional groups, potentially carboxyl groups, or groups containingnitrogen, the predominate species being carbonyl groups. This oxidativefunctionalization would result in water contact angles of 1-75 and atotal carbon to oxygen ratio of less than 2:1.

In certain embodiments, the adherence points would only be imposed uponone side of the para-xylene film. In other embodiments, the adherencepoints are present on both sides of the film. In some embodiments, thefunction of the adherence points would be for the integration orimmobilization of the para-xylene film to the cellular tissue and/ororgan surface or body lining in lieu of using permanent sutures,staples, or some other form of immobilization. Thus, creating anadherent side (oxidized) and non-adherent para-xylene film which wouldserve as a tissue separator and/or adhesion barrier preventinginteraction to said non-adherent side/portion while being anchored(e.g., solely) by adherent points on opposing side.

In certain embodiments, implantation of an adherent/non-adherent filmcan be deployed as a means of ensuring proper wound closure of theintegument (skin), muscle and/or musculature, sub-cutaneous tissue,and/or connective tissue so as to not incur incisional hernias,prevention of incisional hernia after a surgical procedure. Whereas theadherent side (anterior) of the para-xylene film would be affixed to theparietal side of the body cavity and the non-adherent (posterior) sidewould face the visceral side, preventing significant interaction betweenthe tissue and organ surfaces decreasing the likelihood of surgicaladhesions to or around the incision site.

In certain embodiments implantation of adherent/non-adherent film couldbe deployed in an effort to separate individual organ surfaces from oneanother or separate adherence of visceral portions of the peritoneumfrom parietal portions of the body cavities. Placement of anadherent/non-adherent film would be to separate organs from one another,due to the confining nature of the surgical procedure, such para-xylenefilms could be placed between individual organs or folds within theintestines, preventing bowel obstruction, or organ to organ adherence.This situation would present itself, for example, when suturing or othermethods of permanent immobilization cannot be employed due to the lackof space and/or nature of the procedure. In certain embodiments, thepara-xylene films or meshes described herein are used as a trans-vaginalsupport, lift, separator, and/or assist in pelvic organ prolapse, orsupport, lift, and/or separate any other form of organ related prolapsewhich may occur.

In certain embodiments, application of para-xylene film implanted in asubject (e.g., in a temporary fashion) further comprises a tether. Incertain embodiments, the tether is composed of a strand, or elongatedpara-xylene strip originating from the center, end, or some point of thefilm surface and/or edge of said para-xylene film, which uponimplantation the tether/strip would traverse the internal areas ofimplantation and protrude externally through the integument (skin),muscle and/or musculature, sub-cutaneous tissue, and/or connectivetissue at the incision site. Said tether, protruding from the wound siteand/or surgical incision point may, for example, be bandaged over toassure risk of infection is minimized. The presence of the tether and/orstrip would allow for complete removal of para-xylene film after aperiod determined by a clinician, surgeon etc. Inability of cells andtissue to adhere or interact with the film would permit the tether/stripto prevent full wound closure at the site of incision specifically wherethe tether/strip protrudes from the incision site, allowing for removalof the para-xylene film after any number of days following the surgicalprocedure. Immobilization of said tethered para-xylene film, thesecuring of the main internal portion of the film, could be done usingdegradable sutures or similar resorbable means. Upon degradation ofresorbable fixation, the film could/would be safely removed.Manufacturing of temporary para-xylene films with integratedtether/strip could be manufactured, for example, as one piece.

In certain embodiments, deposition of a para-xylene polymer onto theouter surface of a surgical implant that is permanent, non-resorbable,non-degradable mesh or other related permanent implant, is in a mannerthat said polymer uniformly coats the mesh or implant. In particularembodiments, each side of the mesh or implant may be characterized by avisceral side (e.g., typically posterior orientation) and a parietalside (e.g., typically anterior orientation). In certain embodiments,composition of the top surface (bottom surface would adherent to mesh ordevice) where anterior surface is tuned through some method of oxidation(e.g., plasma oxidation and/or UV irradiation) such that the anteriorsurface acts to integrate or promote tissue adherence or attachment,while the non-oxidized posterior surface prevents such cell/tissueinteraction or attachment. Thus, the tuning of the surface will promotedevice integration while the opposing side would prevent suchinteraction to body tissues or organ surfaces either adjacent or thosewhich come into contact with said device. In particular embodiments,this would be especially important for implanted devices within variousbody cavities whereby a surface which could promote integration on onesurface, while preventing or repel tissue attachment to the body cavityor adjacent organ(s) further aiding in preventing adhesions or scarformation to the opposing or opposite non-oxidized surface.

In certain embodiments, the purpose of said coating would be to prevent,resist, and/or repel tissue and/or organ adhesion and/or integration ofsaid implant into the surrounding tissue surfaces and/or organ surfaces,specifically interaction with the visceral peritoneum or the visceralside, side facing or interacting with the organ surface(s) most likelyposterior. This portion of most traditional mesh designs is composed ofa smooth, or less rough side, characterized by a smooth web, netting, orinterweaving of mesh strands created to present a smooth surface meantto face, or come into contact with the visceral peritoneum and/orvisceral organ surfaces. In certain embodiments, such surgical meshdesigns may or may not be composed of parylene as described herein.

In additional embodiments, the parylene film will act (e.g., alone) as ahernia mesh material, or wound closure mesh/material. In certainembodiments, the parylene film possesses perforations, holes, and/orcut-outs (e.g., placed strategically across the film giving theappearance of a mesh, net, and/or webbing). Such an embodiment is shownin FIG. 8. In particular embodiments, the perforations, holes, orcut-out portions within the film allow flexibility and stretching inorder to accommodate the flexing and stretching of integument (skin),muscle and/or musculature, sub-cutaneous, and/or connective tissue whenthe film is implanted. In particular embodiments, the parylene mesh,net, or webbing will have thickness measurements between 1 μm to 5 mm.The thickness of said parylene film will facilitate the strength andflexibility required for a device implanted in such a manner.

In certain embodiments, the para-xylene coating of the parietal facingportion of the surgical mesh is characterized by a rough web, netting,or interweaving of mesh strands creating varying topography such as tomaximize mesh integration into the parietal peritoneum or parietalsurface most likely anterior. The mesh design may or may not be composedof para-xylene as described herein. In certain embodiments, theperitoneal side may be treated to elicit and/or promote interaction,adherence to this surface/side. Oxidation of only this portion or sideof coating/device (anterior) would create adherence, through any meansof oxidation, oxygen plasma etching, etc. In certain embodiments, stepscould be taken, such as through immersing or shielding of the visceralportion, through use of a Polydimethylsiloxane (PDMS) stamp or othermaterial, which could sufficiently prevent oxidation of the visceralside, allowing for oxidation of the parietal surface alone andsubsequent alteration to surface chemistry resulting in a carbon tooxygen content ratios of less than 2:1, contain predominately carbonylgroups, a lesser degree carboxyl groups, but also potentially contain arange of nitrogen groups resulting in water contact angles 1-75,attributed to oxidized para-xylene physical and chemicalcharacteristics, excluding surface roughness which would be determinedby the surface of said mesh.

In particular embodiments, the internal region of the subject isselected from the group consisting of: dorsal cavity, ventral cavity,thoracic cavity, pleural cavity, mediastinum, abdominopelvic cavity,abdominal cavity, pelvic cavity, lumbar region, umbilical region,inguinal region, hypogastric region, abdominal region, peritonealsurfaces, visceral surfaces, mesothelial surfaces, antebrachial,axillary, brachial, buccal, carpal, cervical, coxal, crural, cubital,femoral, mental region, orbital region, patellar region, pubic region,tarsal region, thoracic cavity, gluteal region, lumbar region, occipitalregion, popliteal region, scapular region, sural region, nasal region,cephalic region, oral cavity, otic region, acromial region, deltoidregion, digital area, sternal region, pectoral region, mammary region,pedal region, plantar region, calcaneal region, sacral region, vertebralregion, sagittal region, coronal region, transverse region, obliqueregion, brain, spinal cord, heart, right lung, left lung, right kidney,left kidney, and bladder. In other embodiments, the first area comprisesthe heart and the second area comprises the sternum.

In some embodiments, the present invention provides system comprising:a) an article of manufacture consisting of, or consisting essentiallyof: a polymer film and/or coating with a top surface and a bottomsurface, wherein the polymer film consists of, or consists essentiallyof, a para-xylene polymer(s), wherein the polymer film and/or coatinghas a thickness between 1 μm and 75 μm (e.g., 1 . . . 5 . . . 15 . . .25 . . . 38 . . . 55 . . . or 75 μm), wherein the top surface has awater contact angle between 75 and 95 degrees (e.g., 75 . . . 79 . . .81 . . . 84 . . . 88 . . . 91 . . . or 95 degrees), and wherein thepolymer film is sized to serve as a tissue separator, tissue adherentplatform, and/or adhesion barrier in a subject; and b) a packagingcomponent, wherein the article of manufacture is located inside thepackaging component. In certain embodiments, the article of manufactureis sealed inside the packaging component such that the article ofmanufacture remains sterile while inside the packaging component.

In further embodiments, the present invention provides articles ofmanufacture comprising: a) a device comprising at least one surface, andb) a polymer film coating at least a portion of the at least one surfaceof the device (e.g., medical device), wherein the polymer film has anexposed top surface, wherein the polymer film consists of, or consistsessentially of, a para-xylene polymer (s), wherein the polymer film hasa thickness between 1 μm and 75 μm, and wherein the top surface has awater contact angle between 75 and 95 degrees. In certain embodiments,the device is a medical device selected from the group consisting of: acatheter, a hernia mesh, a stent, an implantable cardioverterdefibrillators (ICD), a balloon dilator, a band ligator, a surgicalclip, forceps, a surgical guidewire, a surgical snare, and animplantable pulse generator.

In particular embodiments, the polymer film is sterilized. In otherembodiments, the para-xylene polymer comprises parylene C. In furtherembodiments, the para-xylene polymer is selected from the groupconsisting of: parylene A, parylene AM, parylene AF4, parylene SF,parylene HT, parylene X, parylene N, and parylene D. In additionalembodiments, the article of manufacture consists of the polymer filmand/or coating. In other embodiments, the subject is a human, cat, dog,horse, cow, pig, or other domesticated animal.

In certain embodiments, the water contact angle for the top surface isbetween 80 and 90 degrees (e.g., about 80, 81, 82, 83, 84, 85, 86, 87,88, 89, or 90 degrees). In further embodiments, the bottom surface has awater contact angle between 75 and 95 degrees (e.g., 75 . . . 79 . . .81 . . . 84 . . . 88 . . . 91 . . . or 95 degrees). In otherembodiments, the bottom surface has a water contact angle of 1-75degrees (e.g., 75 . . . 79 . . . 81 . . . 84 . . . 88 . . . 91 . . . or95 degrees) or 96-180 degrees (e.g., 96 . . . 110 . . . 120 . . . 150 .. . or 180). In certain embodiments, the bottom surface is oxidized. Insome embodiments, the polymer film has a thickness between 5 μm and 25μm (e.g., 5 . . . 10 . . . 15 . . . 20 . . . or 25 μm). In furtherembodiments, the top surface has a RMS roughness between 1.0 nm and 10.0nm (e.g., 2.5 . . . 3.5 . . . 5.0 . . . 6.5 . . . or 7.5 nm). Inadditional embodiments, the bottom surface has a RMS roughness between1.0 nm and 10.0 nm (e.g., 2.5 . . . 3.5 . . . 5.0 . . . 6.5 . . . or 7.5nm). In other embodiments the film has an optical transmission of60-99.9% in the high visible (400-1000nm) and near infra-red (3200-6000cm) range.

In some embodiments, the present invention provides compound films. Forexample, in certain embodiments, the present invention provides articlesof manufacture comprising, consisting essentially of, or consisting of:a compound film, wherein the compound film comprises, consistsessentially of, or consists of: a) a base polymer film with a base topsurface and a base bottom surface, wherein the base polymer filmcomprises a first para-xylene polymer and has a thickness between 1 μmand 75 μm (e.g., 1 . . . 5 . . . 15 . . . 25 . . . 38 . . . 55 . . . or75 μm); b) a first therapeutic molecule layer disposed on the base topsurface, wherein the first therapeutic molecule layer comprises firsttherapeutic molecules; and c) a first cover polymer film with a firstcover top surface and first cover bottom surface, wherein the firstcover bottom surface covers the first therapeutic molecule layer, andwherein the first cover polymer film comprises a second para-xylenepolymer and has: i) an elution-allowing (sub-conformal) thicknessbetween about 150 nm and 600 nm (e.g., 150 . . . 250 . . . 350 . . . 450. . . 550 . . . and 600 nm) mediating passive transport of thetherapeutic(s) across said polymer film cover, or ii) a non-elutionallowing (conformal) thickness between about 1 μm and 75 μm (e.g., 1 . .. 5 . . . 15 . . . 25 . . . 38 . . . 55 . . . or 75 μm).

In certain embodiments, the present invention provides methods forseparating areas in an internal region of a subject (and/or treatinginflammation or atrial fibrillation) comprising: inserting an article ofmanufacture as described herein (e.g., containing a compound film) intoan internal region of a subject between first and second areas. Inparticular embodiments, the internal region of the subject is selectedfrom the group consisting of: dorsal cavity, ventral cavity, thoraciccavity, pleural cavity, mediastinum, abdominopelvic cavity, abdominalcavity, pelvic cavity, lumbar region, umbilical region, inguinal region,hypogastric region, abdominal region, peritoneal surfaces, visceralsurfaces, mesothelial surfaces, antebrachial, axillary, brachial,buccal, carpal, cervical, coxal, crural, cubital, femoral, mentalregion, orbital region, patellar region, pubic region, tarsal region,thoracic cavity, gluteal region, lumbar region, occipital region,popliteal region, scapular region, sural region, nasal region, cephalicregion, oral cavity, otic region, acromial region, deltoid region,digital area, sternal region, pectoral region, mammary region, pedalregion, plantar region, calcaneal region, sacral region, vertebralregion, sagittal region, coronal region, transverse region, obliqueregion, brain, spinal cord, heart, right lung, left lung, right kidney,left kidney, and bladder. In certain embodiments, the first areacomprises the heart and the second area comprises the sternum. Inadditional embodiments, the first or second cover polymer film has anelution-allowing thickness between 150 nm and 600 nm (e.g., 150 . . .250 . . . 350 . . . 450 . . . 550 . . . and 600 nm) mediating passivetransport of the therapeutic(s) across said polymer film cover, andwherein the first and/or second therapeutic molecules pass out of thecompound film into the internal region of the subject. In otherembodiments, the first or second cover polymer film has a non-elutionallowing thickness between 1 μm and 75 μm (e.g., 1 . . . 5 . . . 15 . .. 25 . . . 38 . . . 55 . . . or 75 μm), and wherein the method furthercomprises suturing the compound film in the internal region in thesubject, wherein the suturing generates holes in the compound film thatallow the first and/or second therapeutic molecules to pass out of thecompound film into the internal region of the subject.

In certain embodiments, the present invention provides systemscomprising: a) an article of manufacture as described herein (e.g., witha compound film); and b) a packaging component, wherein the article ofmanufacture is located inside the packaging component. In particularembodiments, the article of manufacture is sealed inside the packagingcomponent such that the article of manufacture remains sterile whileinside the packaging component. In other embodiments, the compound filmhas a thickness (i.e., total thickness) between about 1 μm and 75 μm. Inother embodiments, the article of manufacture further comprises adevice, wherein at least a portion of the device is coated with thecompound film. In additional embodiments, the device comprises a medicaldevice.

In further embodiments, the first cover top surface is an outermostlayer of the compound film and has a water contact angle between 75 and95 degrees. In other embodiments, the compound film further comprises:d) a second therapeutic molecule layer (and optionally a third, fourth,fifth, sixth, etc. therapeutic molecule layer) disposed on the firstcover top surface (and disposed on the next available outer surface forthe third, forth, fifth, sixth, etc. therapeutic molecule layers),wherein the second therapeutic molecule layer comprises secondtherapeutic molecules (and the additional layers comprise third, fourth,fifth, etc. therapeutic molecule layers). In some embodiments, thecompound film further comprises: e) a second cover polymer film with asecond cover top surface and a second cover bottom surface, wherein thesecond cover bottom surface covers the second therapeutic moleculelayer, and wherein the second cover polymer film comprises a thirdpara-xylene polymer and has: i) an elution-allowing thickness betweenabout 150 nm and 600 nm, or ii) a non-elution allowing thickness betweenabout 1 μm and 75 μm.

In particular embodiments, the second cover top surface is an outermostlayer of the compound film and has a water contact angle between 75 and95 degrees. In further embodiments, the article of manufacture consistsof, or consists essentially of, the compound film, and wherein thecompound film is sized to serve as a tissue separator and/or adhesionbarrier in a subject. In other embodiments, the base bottom surface isan outermost layer of the compound film and has a water contact anglebetween 75 and 95 degrees.

In other embodiments, the compound film further comprises: d) a secondtherapeutic molecule layer disposed on the base bottom surface, whereinthe second therapeutic molecule layer comprises second therapeuticmolecules. In other embodiments, the compound film further comprises: e)a second cover polymer film with a second cover top surface and a secondcover bottom surface, wherein the second cover bottom surface covers thesecond therapeutic molecule layer, and wherein the second cover polymerfilm comprises a third para-xylene polymer and has: i) anelution-allowing thickness between about 150 nm and 600 nm, or ii) anon-elution allowing thickness between about 1 μm and 75 μm or 1 um and5 mm.

In certain embodiments, in lieu of a therapeutic being deposited orintegrated between 2 conformal or encapsulating layers or para-xylene,the integration of a wireless device/sensor is employed. This wirelessdevice or sensor could be used, for example, to track certain patientenvironmental or physical conditions, where said sensor would then relaypatient related information wirelessly when a receiver or recorder isplaced on or near the implant and/or patient. The sensor would besandwiched or encapsulated between para-xylene layers. Manufacture ofthe sensor could be achieved by a single thinner deposition ofpara-xylene, placing or laying of the wireless device or sensor and thendepositing another thicker layer on top of the initial para-xylene layerand device. Resultant device would be a thin base layer and then asignificantly thicker second layer bonding or sealing device between twolayers of para-xylene with a device or sensor in the middle between thetop or bottom surface.

In certain embodiments, the compound film is sterilized. In furtherembodiments, the first and/or second para-xylene polymer comprisesparylene C. In other embodiments, the first and/or second (and/or third,fourth, etc.) para-xylene polymer is selected from the group consistingof: parylene A, parylene AM, parylene AF4, parylene SF, parylene HT,parylene X, parylene N, and parylene D. In some embodiments, the basetop surface is oxidized. In additional embodiments, the base polymerfilm has a thickness between 5 μm and 25 μm (e.g., 5 . . . 10 . . . 15 .. . 20 . . . or 25 μm). In additional embodiments, the first cover topsurface has a RMS roughness between 1.0 nm and 10.0 nm (e.g., 2.5 . . .3.3 . . . 4.1 . . . 6.3 . . . or 7.5 nm). In other embodiments, the basebottom surface has a RMS roughness between 1.0 nm and 10.0 nm (e.g., 2.5. . . 3.3 . . . 4.1 . . . 6.3 . . . or 7.5 nm). In further embodimentsthe film has, an optical transmission of 60-99.9% in the high visible(400-1000 nm) and near infra-red (3200-6000 cm) range.

In certain embodiments, the size and shape of the film will take avariety of forms. This includes square to rectangular forms with roundededges (e.g., 2″×3″, 3″×3″ 3″×4″, 5″×6″, 6″×6″, 6″×7″ inches, etc)thereby revealing a shape with no sharp corners and/or edges. In furtherembodiments the shape will take a circular (e.g., as shown in FIG. 7),oval, or ovoid shapes or form (e.g., 2″, 3″, 4″ 5″ inch diameters, etc).Other embodiments may take the form of a triangular shape, possessingrounded edges (e.g., 2″, 3″, 4″ 5″ inch heights, etc.). In additionalembodiments, the film will be provided in a larger single piece (e.g.,9″×9″, 9″×10″, 10″×11″ inches, etc) where the end user, surgeon, and/orclinician will cut the film to shape according to procedure and/orapplication, or intended use. In other embodiments, the film will have aclover leaf, or club shape or appearance to include a two, three, orfour leaf clover/club type design to facilitate the wrapping or abilityto conform to spherical or three dimensional shaped objects such asvarious body organs and/or body cavities without excessive bunching,wrinkling or crimpling along certain edges or portions of said film.

DESCRIPTION OF THE FIGURES

FIGS. 1A-B: A) Schematic representation of an exemplary stand aloneParylene-C (PPX) derived Drug Delivery Platform (DDP) (e.g., forimplantation to reduce incidence of perioperative inflammation andpostoperative atrial fibrillation). The transparency and thin profile ofthe film are physical properties lending to the amenable nature of thedevice. Integration of therapeutics, such as Dexamethsaone (DEX) andAmiodarone (AMIO), into the PPX film increases the effectiveness withwhich the device can counteract complications stemming from surgery(e.g., cardiovascular surgery). B) Postoperative images taken at thetime of repeat sternotomy showing film involvement among theexperimental conditions as described in Example 1 below.

FIGS. 2A-F: Quantitative assessment of RAW264.7 macrophages on varyingsurfaces to determine cellular response as described in Example 1 below.A) Cellular viability of macrophages grown on Control and PPX surfacesover the respective time points. Further assays indicate B)proliferative ability of macrophages grown over time periods shown aswell as an adhesion assay. Cellular interaction via an adhesion assay C)to PPX surfaces was performed with bare surfaces (−Cells) in addition tosurfaces with a confluent monolayer of cells present (+Cells).Quantitative assessment of 3T3-NIH fibroblasts on varying surfaces todetermine cellular response. D) Cellular viability of fibroblasts grownon Control and PPX surfaces over the respective time points. Furtherassays indicate E) proliferative ability of fibroblasts grown over timeperiods shown as well as an adhesion assay. Cellular interaction via anadhesion assay F) to PPX surfaces was performed with bare surfaces(−Cells) in addition to surfaces with a confluent monolayer of cellspresent (+Cells.) *P<0.001, †P<0.01, §P<0.05, ¥ P<0.005

FIGS. 3A-E: RAW264.7 macrophage expression of IL-6 A) in response to PPXsurfaces under examination as described in Example 1 below. Tissueculture plates (Control) were coated with PPX, modified accordingly(PPX-Oxd, PPX and PPX•) and then seeded with RAW264.7 macrophages.Baseline expression of IL-6 reveals modified and unmodified PPX surfacesdo not increase IL-6 cytokine expression. Intentional inflammatorystimulation (+), with LPS, was completed to emphasize therapeuticresponse. Suppression of stimulated inflammation through DEX integrationshows capacity of PPX film to locally deliver therapeutics. *P<0.001,†P<0.01, §P<0.05 Cross section images B) reveal relative surfacewettability of each substrate. Angles obtained from image C)measurements show wettability characteristics of each substrate.Parylene (PPX) modified surfaces provide a contrasting effect of plasmaoxidation on non-oxidized (PPX) and oxidized parylene (PPX-Oxd).Oxidation of PPX surfaces mimics wettability content of Controlsurfaces. Current surgical films (Repel™ and Preclude™) show contrastinglevels of surface wettability. Surface feature measurements D) completedvia AFM reveal RMS roughness of film surfaces compared to the Controlsurface. PPX films present a surface that is less rough when compared toRepel™ or Preclude™ which presents a relatively rough surface tocellular tissues. Further profile measurements E) which identify thethickness of each of the films examined, indicate PPX films provide athinner profile when compared to surgical films Repel™ and Preclude™.*PPX deposition is 40 grams of Parylene-C

FIGS. 4A-J: X-Ray Photoelectron Spectroscopy (XPS) measurementsidentifying constituent elements present on substrate surfaces that wereemployed in Example 1 below. Elemental analysis revealed varying carbonand oxygen ratios as the primary elemental composition on each of thefilms, excluding the Preclude™ membrane. Control surfaces (A) revealed agreater degree of carbon to oxygen content. Comparatively, PPX (B)showed a decrease in oxygen content when compared to available carbon.Oxidized PPX (C) (PPX-Oxd) reveals an increase in oxygen which can beattributed to the oxygen plasma surface bombardment. It is of interestto note, the carbon/oxygen balance similarities between Control andPPX-Oxd substrates. The Repel™ (D) film is the only surface presentwhose oxygen content supersedes the amount of carbon present. Preclude™(E) remaining the striking exception, as it contains a predominateamount of fluorine on the surface with negligible carbon content. FT-IRsurface scans representing chemical species present on the surface ofthe respective films. F) Control surfaces, consisting of plasma oxidizedpolystyrene compared to various PPX conditions. G) Native PPX surfaceand H) oxidized PPX (PPX-Oxd) appear quite similar other than slightalterations in and around the 3000 cm⁻¹ peak. Repel™ I) and Preclude™ J)spectra are provided for comparison of current clinical anti-adhesivebarriers.

FIGS. 5A-D: This figure presents in vivo results from Example 1 below.The extent of perioperative inflammation represented as adhesion scoresA) to implanted PPX films. 3 point scoring system depicting level ofinflammation and scarring which occurred, 0: No Adhesions, 1: FilmyAdhesions, 2: Moderate Adhesions 3: Severe Adhesions. Therapeuticinvolvement decreased inflammation related adhesions considerably(PPX-Oxd[DEX] and PPX[AMIO, DEX] films compared to Control. Native PPXfilms without drug prevented adhesion formation due to inherent materialproperties. Alteration of PPX films through surface oxidation (PPX-Oxd)incited severe inflammation as noted by an increase in adhesionformation. *P<0.001, †P<0.01 Tissue section staining B) reveal level ofscarring and cellular infiltration, respectively. Level of sustainedinflammation is apparent in Control and PPX-Oxd samples from the extentof NTF formation and cell density. Reduced infiltrates are noted inPPX-Oxd[DEX] and PPX[AMIO, DEX] samples a result of locally deliveredDEX. PPX alone displayed cellular infiltrates greater than DEX infusedfilms, but less than Control or modified PPX surfaces. Tissue analysisC) reveals NTF thickness as a result of perioperative inflammation.Thickness measurements of NTF correlate to levels of inflammation at thetime of resternotomy. PPX films integrated with DEX showed less NTFcompared to Control and PPX-Oxd films. Contrastingly, PPX alone showedfibrosis thickness similar to that of DEX infused films. *P<0.001 Atrialfibrillation experiments (D) underscore the ability for a PPX film toreduce entrance into and duration of AF when loaded with AMIO anddelivered locally. §P<0.05

FIGS. 6A-C: Time lapse images of RAW264.7 murine macrophages A) grown onvarying substrate surfaces over the course of 72 hrs (images taken atcorresponding time points 4, 24, 48 and 72 hrs) as described inExample 1. As indicated by the Control surface RAW264.7 macrophagesdisplay comparative monolayer outgrowth on either of the oxidized PPXsurfaces (PPX-Oxd, PPX-Oxd•). A somewhat irregular growth pattern isseen on PPX surfaces which did not undergo plasma oxidation. RAW264.7macrophages were seeded at appropriate confluency to allow for maximumoutgrowth on the third day. Cells were grown under standard conditionshumidified air maintained at 37° C., 5% CO² supplemented with DMEM 10%FBS, 1% Pen/Strep. Time lapse images of 3T3-NIH murine fibroblasts B)grown on varying substrate surfaces over the course of 72 hrs (imagestaken at corresponding time points 4, 24, 48 and 72 hrs). As indicatedby the Control surface 3T3-NIH fibroblasts display comparative monolayeroutgrowth on either of the oxidized PPX surfaces (PPX-Oxd, PPX-Oxd•).Striking irregular growth patterns are evident on PPX surfaces which didnot undergo plasma oxidation, lending to the assertion the fibroblastswere unable to adhere to the PPX surface. 3T3-NIH fibroblasts wereseeded at appropriate confluency to allow for maximum outgrowth on thethird day. Cells were grown under standard conditions humidified airmaintained at 37° C., 5% CO² supplemented with DMEM 10% FBS, 1%Pen/Strep. Atomic Force Microscopy (AFM) images C) of surfaces (Control,PPX and PPX-Oxd) and films (Repel™ and Preclude™) under investigation.Image sets denoted by A & C reveal phase images compared totopographical outputs on B & D. Phase images reveal similar materialproperties across the surface. Uniform shading indicates the surface iscomposed of a similar material. Topographical images reflect changes inelevation across the film surface. Alterations in shading correspond todepressions or elevations across the film surface. Images A & B and C &D represent zoom in and zoom out functions respectively.

FIG. 7: shows an exemplary circular embodiment of the film of thepresent invention.

FIG. 8: shows an exemplary circular embodiment of the film of thepresent invention with various perforations, holes, and/or cut-outs.

DETAILED DESCRIPTION

The present invention provides single sheet and compound para-xylenefilms for therapeutic uses. For example, the present invention providessingle sheet para-xylene films useful as tissue separators and/oradhesion barriers in a subject, where the top and/or bottom surfaces ofsuch films have a water contact angle between 75 and 95 degrees (e.g.,to prevent adhesion formation). The present invention also providescompound films composed of at least two para-xylene polymer films with atherapeutic molecule layer in between. Such compound films, when used invivo (e.g., as a tissue separator and to treat inflammation or atrialfibrillation) allow either therapeutic molecule elution through one ofthe para-xylene layers, or therapeutic molecule release when thecompound film is pierced, such as when it is sutured in place.

The present invention is not limited by the size of the single sheet andcompound films. Preferably, these films are sized to fit inside asubject to serve as a tissue separator and/or adhesion barrier. Incertain embodiments, the films are 18×13.5 cm; or 9×9 cm; or 10×10; or20×7 cm. In certain embodiments, the surface area of the films is about200 cm²; or between 10 and 400 cm² (e.g., 10 . . . 30 . . . 75 . . . 140. . . 243 . . . 333 . . . 380 . . . or 400 cm²). The present inventionis not limited by the shape of the films. Exemplary shapes includecircles, ovals, squares, octagons, etc.

The present invention is not limited by the type of therapeuticmolecules that can be included in the compound films. Therapeuticmolecules include, but are not limited to, any type of biologics,dexamethasone, amiodarone, biosimilars, thrombin inhibitors,antithrombogenic agents, thrombolytic agents, fibrinolytic agents,vasospasm inhibitors, calcium channel blockers, vasodilators,antihypertensive agents, antimicrobial agents, antibiotics, inhibitorsof surface glycoprotein receptors, antiplatelet agents, antimitotics,microtubule inhibitors, anti-secretory agents, actin inhibitors,remodeling inhibitors, antisense nucleotides, anti-metabolites,anti-proliferatives, anticancer chemotherapeutic agents,anti-inflammatory steroid or non-steroidal anti-inflammatory agents,immunosuppressive agents, growth hormone antagonists, growth factors,dopamine agonists, radiotherapeutic agents, extracellular matrixcomponents, inhibitors, free radical scavengers, chelators,antioxidants, anti-polymerases, antiviral agents, photodynamic therapyagents, gene therapy agents; small molecules, proteins, multiproteinmacromolecules (e.g., antibodies), nucleic acids (including, but notlimited to, siRNA, shRNA, miRNA, etc.); hydrophilic small moleculedrugs, hydrophobic small molecule drugs, steroidal small molecule drugs,macrocyclic small molecule drugs, small molecule drugs without bulkyside groups, small molecule drugs with bulky side groups, small moleculedrugs in pharmaceutical acceptable salt forms, peptide biologics,protein biologics, multi-chain protein biologics, glycosylated proteinbiologics, immunoglobulins, micro chain nucleic acid biologics, shortchain nucleic acid biologics, nucleic acid biologics, aptamer biologics,protein-nucleic acid complex biologics, lipid biologics, lyposomebiologics and PEGylated forms of any of the foregoing, dexamethasone,doxorubicin, IgG, interferon2b, mitomycin, clopidogrel, paclitaxel,hormones, hormone mimetics and hormone derivatives, including planthormones.

In certain embodiments, the films of the present invention are providedin a system or kit along with a packaging component (e.g., container forthe films when they are stand-alone components for tissue separation oradhesion barriers). In certain embodiments, the packaging component issized to fit the film and is a foil lined bag commonly used for medicaldevices, such as adhesion barriers. In particular embodiments, thepackaging component is a gas-impermeable, foil bag. In some embodiments,the packaging component further includes written instructions for propercare and/or use of the films.

The present invention is not limited by the fabrication technique usedto make the films of the present invention. For example, the films canbe fabricated upon a solid surface as a temporary platform which can beeventually removed for standalone activity. This solid surface can be,for example, a glass slide, coverslip, silicon wafer, plastic disc andthe like. In certain embodiments, the films can be generated as follows.Onto to a solid surface, a base layer of para-xylylene polymer isdeposited. This can be done, for example, via a room temperaturechemical vapor deposition process. In the chemical vapor depositionprocess, the para-xylene monomer is sprayed as a monolayer onto thesolid surface such that the monomer reversibly attaches to the solidsurface and self-polymerizes. The result being a film of para-xylyleneis created by deposition onto the solid surface. In certain embodiments,a Specialty Coating Systems Lab Coater, per the manufacturer's protocol,can be used to deposit a base layer of pary-xylene (e.g., parylene C)onto a glass disc via a room temperature chemical vapor deposition.

The surface of the films of the present invention, in some embodiments,may be oxidized. Oxidative functionalization may be accomplished, forexample, by ultraviolet light, a plasma cleaner, chemically drivenoxidization or any other oxidation processing of the para-xylene.

In certain embodiments, a layer of therapeutic molecules is depositedonto a surface of the films of the present invention. The therapeuticmay be, for example, physically, ionically, or covalently linked to thesurface of the layer. An exemplary method to dispose about, permeateand/or deposit the therapeutic on or to a layer is a spotting followedby evaporation. In this method, a solvent containing the therapeutic isdeposited on the layer. The solvent then evaporates off slowly. Assolvent evaporates off, the therapeutic falls down and depending uponthe wettability of the layer, the deposited therapeutic diffuses in thelayer. This leaves a dry therapeutic that is permeated in and/ordisposed about the surface of the layer and forms a reservoir oftherapeutic.

In certain embodiments, a plurality of distinct laminated layers ormultilayer can e deposited by repeating the above process. In subsequentchemical vapor depositions, the para-xylene monomer is sprayed onto anunderlying layer of polymerized para-xylylene. Similar to when themonomer is sprayed on the solid surface, it self polymerizes. Unlikewhen the monomer is sprayed on the solid surface, when sprayed on anunderlying layer of polymerized para-xylylene, it essentiallyirreversibly attaches to form a laminate. This process is repeated asmany times to build up the number of desired layers. In betweenrepeating the deposition process, an oxidative functionalization processcan be performed.

EXAMPLES Example 1 Para-Xylene Polymer Films for Adhesion Prevention andReduction in Inflammation

This Example describes the construction and use of para-xylene polymerfilms to prevent adhesions and inflammation in vivo.

Suppression of both perioperative inflammation and post-operative atrialfibrillation has been hindered due to an absence of effective drugdelivery platforms (DDP) in the post-operative cardiac setting.Localized released of an anti-inflammatory and anti-arrhythmic agentsmay be more effective than intravenous drug delivery in order to improvepatient outcomes. An investigation utilizing a Parylene-Cnano-structured therapeutic film infused with dexamethasone (DEX) andamiodarone (AMIO) to inhibit inflammation and atrial fibrillation wasperformed as described below.

A Parylene-C (PPX) film was tested in an established rabbit model ofpericardial adhesion formation. Following sternotomy, the anteriorpericardium was resected and the epicardium was abraded. Rabbits wererandomly assigned to five treatment groups: Control (plasma oxidizedpolystyrene), oxidized PPX (PPX-Oxd), PPX-Oxd infused with DEX(PPX-Oxd[DEX]), native PPX (PPX), and PPX infused with DEX and AMIO(PPX[AMIO, DEX]). After 4 weeks post-sternotomy, pericardial adhesionswere evaluated for gross adhesions using an established 4-point gradingsystem, histological evaluation for neotissue fibrosis (NTF) on theepicardium. Atrial fibrillation duration and time per induction weremeasured.

The PPX[AMIO, DEX] group had a significant reduction in mean adhesionscore compared with the Control group (Control 2.75±0.42 vs. PPX[AMIO,DEX] 0.25±0.42) (P<0.01). The PPX[AMIO. DEX] group was similar to nativePPX (PPX 0.38±0.48 vs. PPX[AMIO, DEX] 0.25±0.42) (P=NS). Adhesions inthe PPX-Oxd group were not distinguishable from the Control group(PPX-Oxd 2.83±0.41 vs Control 2.75±0.42) (P=NS). A reduction inneotissue fibrosis was present in the PPX[AMIO, DEX] group compared tothe Control group (PPX[AMIO, DEX]0.80±0.10 mm vs. Control 1.78±0.13mm)(P<001). Total duration of atrial fibrillation was decreased inrabbits with PPX[AMIO, DEX] films compared to Control (9.5±6.8 sec vs.187.6±174.7 sec, p=0.003). Time of atrial fibrillation per successfulinduction decreased among PPX[AMIO, DEX] films compared to Control(2.8±1.2 sec vs. 103.2±178 sec, p=0.004).

These results indicated that DEX and AMIO impregnated Parylene-C filmsare associated with reduced perioperative inflammation and a diminishedduration of atrial fibrillation. Epicardial application of AMIO, DEXfilms is a useful strategy to prevent post-operative cardiaccomplications.

Materials and Methods Material Fabrication

20 grams of Parylene-C (dichloro(2,2)paracyclophane) (PPX) (ParatechCoating Inc., Aliso Viejo, Calif.) was deposited onto SSP Type 110silicon wafers (University Wafer, Boston, Mass.). Deposition occurredwithin a Labcoter 2 PDS 2010 (Specialty Coating Systems SCS,Indianapolis, Ind.) under default conditions.¹³ Drug loading involvedapplying 150 μg of DEX (Alfa Aesar, Ward Hill, Mass.) and 800 μg of AMIO(U.S. Pharmacopeia, Rockville, Md.) on PPX surfaces (600 μg of DEX and3.2 mg of AMIO total). Following drug loading, a 0.4 gram sub-conformalPPX layer was deposited on the drug layer followed by plasma oxidationof the surface. A second drug layer was then applied, and a second 0.4gram sub-conformal PPX was deposited on top of the drug layer. This lastlayer was not oxidized and instead was left hydrophobic. Sterilizationof drug loaded films was completed through exposure to ethylene oxidegas utilizing an Anprolene AN74i gas sterilizer (Anderson Products, Inc.Haw River, N.C.) per manufacturer's recommendations.

In-vitro Analysis

In-vitro analysis and drug release was completed through deposition ofPPX on varying culture plates. Surface treatment of prepared PPX cultureplates either had plasma oxidation (Oxd) or lack thereof resulting inhydrophilic or hydrophobic surfaces (native form). RAW264.7 macrophagesand NIH-3T3 fibroblasts were analyzed on varying surfaces to determinein-vitro response. Cells were kept in DMEM media supplemented with 10%FBS and 1% Pen/Strep in humidified atmosphere with 5% CO². Cellularanalysis was completed through the application of time lapse brightfield imaging in addition to multiple cellular assays examiningviability (MTT, Roche Diagnostics), proliferation (CyQUANT, LifeTechnologies) and adhesion (Vybrant, Molecular Probes) onto polystyrene,representing baseline biocompatible control surfaces, and various PPXsurfaces. RNA isolation was accomplished utilizing TRIzol reagent(Invitrogen Corporation, Carlsbad, Calif.) per the manufacturer'sguidelines. cDNA was synthesized using the iScript Select cDNA SynthesisKit (Quanta Biosciences, Gaithersburg, Md.). PCR was done using SYBERGreen detection reagents (Quanta Biosciences, Gaithersburg, Md.) andappropriate primers for IL-6 and (3-Actin (Integrated DNA Technologies,Coralville, Iowa). Samples were amplified using a MyiQ real-time PCRdetection system (Bio-Rad).

Material Characterization

X-ray Photoelectron Spectroscopy (XPS) was completed using an OmicronESCA probe (Omicron NanoTechnology, Eden Prairie, Minn.) coupled with anEA125 hemispherical energy analyzer. Photoemission was stimulatedthrough monochromated Al (KR) radiation (1486.6 eV) with a power outputof 300 W under ultra-high vacuum (UHV). Survey scans completed by theanalyzer were maintained in constant analyzer energy (CAE) mode at 50eV. Binding energies were referenced at the C is (285.0 eV) bindingenergy set. The binding energy spectrum for each XPS survey scan rangedfrom 0-1200 eV. Surface wettability was determined through staticcontact angle measurements using a VCA Optima contact angle goniometer(AST Products, Inc., Billerica, Mass.) equipped with an automatedpipetting system. Ultrapure H₂O volumes of 15 μl were dispensed onvarying substrates and subsequent images and angle measurements werecollected through AutoFAST Imaging and SPC software respectively. Anglemeasurements were collected in air and under ambient temperatures.Atomic Force Microscopy (AFM) images were collected utilizing a CPResearch (Formally ThermoMicroscopes now Veeco Instruments Inc.,Plainview N.Y.) AFM. Imaging was performed in intermittent contact modeusing a Si probe, (μMasch, NSC36A) with a nominal tip radius curvatureof 10 nm. All measurements occurred under atmospheric conditions andambient temperatures. AFM images were rendered and analyzed using WSXMSPM analysis software.¹⁴ Profile measurements concerning film thicknesswere completed utilizing a Veeco Dektak 150 Surface Profiler (VeecoInstruments Inc., Plainview, N.Y.) using a standard scan option equippedwith a 2.5 μm stylus with an applied force of 5.0 mg. Scan length andduration were confined between 1500-2000 μm and 90 seconds respectively,resulting in horizontal resolutions of 0.055 μm to 0.075 μm and verticalresolution maximum of 524 μm. All measurements were completed in air andunder ambient conditions. Analysis was completed using Dektak V9Software. Attenuated total reflection (ATR) Fourier transform infrared(FT-IR) spectroscopy was performed utilizing a Thermo Nicolet Nexus 870IR Spectrophotometer (ThermoFisher Scientific Inc., Waltham, Mass.).FT-IR analyses of the films under investigation were loaded onto anattenuated reflectance kit (ARK) with a Zinc Selenium (ZnSe) crystalcontained within a N2 purged chamber. A liquid nitrogen cooled mercurycadmium telluride (MCT) detector completed 64 scans with a resolution of8 cm⁻¹ over a range of 4,000-650 cm⁻¹ at room temperature. Samples wereloaded onto the ZnSe crystal at a point to point contact interface undermaximum allowable load/pressure. ATR and further baseline correctionswere completed utilizing Omnic and eFTIR software.

Animal Preparation

Thirty New Zealand white female rabbits (4 kg) were randomly assigned toone of five groups: Control group (N=6), oxidized PPX (PPX-Oxd, N=6),PPX-Oxd infused with DEX (PPX-Oxd[DEX],N=6), native PPX (PPX, N=4), andPPX infused with DEX and AMIO (PPX[AMIO, DEX], N=6). Initial sedationwith acepromazine (0.5 mg/kg subcutaneous) was followed by generalanesthesia induction using ketamine (40 mg/kg) and xylazine (7 mg/kg).Endotracheal intubation was achieved with a 3- or 4-mm tube andanesthesia was maintained intraoperatively with 1.5-2.5% isoflurane. Anintravenous catheter was placed in the marginal ear vein.Cardiorespiratory monitoring was maintained throughout surgery.

Rabbit Pericardiotomy Model

Following clipping of the fur at the operative site, the skin wasprepped with betadine. Approach through a midline sternotomy allowedcomplete anterior pericardiotomy to be made between the left and rightphrenic nerves. To incite microvascular bleeding and pericardialadhesion formation, a gauze pad was used to abrade the epicardialsurface of the pericardium for 5 minutes. Experimental biologicmembranes were fixed to the edges of the pericardium using four 5.0polypropylene sutures. Control animals had 4 sutures placed in the openpericardium but no membrane. Tube thoracostomy was employed duringclosure of the sternotomy and subsequently removed.

Arrhythmia Induction and Measurement

Defibrillation patches were placed on the lateral aspect of the thoraxto provide defibrillation if required and to produce a single-leadsurface electrocardiogram. The electrocardiogram was recorded using aMedtronic LIFEPAK® 20/20e Defibrillator/Monitor. The atrium wasidentified by morphology and contraction sequence. Bipolar atrial wireswere affixed (Medtronic temporary myocardial pacing wires, model number6494). A bipolar electrogram was used to confirm placement, if needed. Asurface electrogram was generated at a paper speed of 25 mm/sec and theatrium was exposed to an International Electrotechnical Commission 6LR61alkaline battery to produce atrial fibrillation. Atrial fibrillation wasconfirmed on the surface electrogram, characterized as high-frequency,low-amplitude atrial electrograms, typically with varying R-R intervals.Because not every exposure to current induced atrial fibrillation,duration of exposure was measured for each attempt, measured as thenumber of seconds during which the atrial bipolar wires were exposed tothe induction current. For each successful induction, atrialfibrillation duration was measured as the number of seconds after theend of induction while the atrium remained in fibrillation.

Cardiac Adhesion Assessment

Four weeks following pericardial abrasion and treatment, rabbits againunderwent anesthesia and repeat sternotomy for the assessment of cardiacadhesion formation. Retrosternal scar tissue was observed by a blindedanalyst and adhesion density at the apex, middle and base of the centralepicardium was scored. The gross evaluation was evaluated with a 4 pointscoring system (0-3). The scoring system indicated the following degreeof adhesions 0: No Adhesions, 1: Mild Adhesions (easily dissected), 2:Moderate Adhesions and 3: Severe Adhesions (difficult to dissect). Rightventricular epicardial tissue samples were fixed in 10% formalin,paraffin embedded and hematoxylin-eosin stained for microscopicevaluation of NTF and adhesion formation. Thickness of NTF, as seen withMasson trichome staining, was observed by a blinded analyst with theNational Institutes of Health Image program (version 1.62; NationalInstitutes of Health, Springfield, Va.) to measure cardiac adhesionthickness.

Statistical Methods

All data are presented as mean±standard deviation. Normally-distributeddata are compared by Student t test to analyze variance. Data that werenot normally distributed were analyzed via Wilcoxon rank-sum test. A pvalue of less than 0.05 was considered significant. The primary outcomemeasure for arrhythmia was duration of atrial fibrillation persuccessful induction attempt. In addition, the number of attemptsrequired to put rabbits into atrial fibrillation and duration ofexposure to induction current was tabulated.

Results

An in-depth in-vitro response to PPX surfaces was completed to assesscellular interaction. To identify cellular response to the modified PPXsurfaces, murine derived RAW264.7 macrophages (RAW) and 3T3-NIHfibroblasts (3T3) were seeded onto multi-well tissue growth platespreviously deposited with varying PPX conditions (PPX-Oxd, PPX and PPX•)(•=unloaded PPX mesh deposition). Bright field images taken at timepoints of 4, 24, 48 and 72 hrs reveal the growth pattern of RAW and 3T3cells on corresponding surfaces (FIG. 6A-B). PPX surfaces which had beenmodified via vacuum plasma treatment (PPX-Oxd and PPX-Oxd•) provided aninterface conducive to monolayer cellular outgrowth of both RAW (FIG.6A) and 3T3 cells (FIG. 6B) when compared to control surfaces (oxidizedpolystyrene, tissue culture plastic). Un-modified PPX surfaces (PPX)presented an interface less amenable to normative cellular outgrowth.

Further cellular analysis was completed through the use of viability,proliferative and adhesion assays (FIG. 2). Viability assays werecompleted over the course of 24, 48 and 72 hour time points for eachcell type (FIG. 2A, D). RAW macrophages displayed little variation whenseeded upon various PPX surfaces in reference to control surfaces. Froma proliferative standpoint (FIG. 2B) there is a slight variation as itapplies to the PPX surface, as there is a marginal increase in theproliferative capacity of the macrophages. This variation appears to bedue to the inability of the macrophages to adhere to the surface of thePPX surface as noted by the adhesion assays performed (FIG. 2C).Alternatively, 3T3 fibroblasts revealed similar results as it pertainsto PPX surfaces. Viability (FIG. 2D) was somewhat reduced, specificallyas it applied to PPX surfaces. Combined with the bright field images of3T3 cells (FIG. 6B) on PPX surfaces and the adhesion assays (FIG. 2F)completed, the decreased viability (FIG. 2D) could be the result of pooradhesion to the surface. Despite relative viability values comparable tocontrols, the proliferative capacity of 3T3 cells was reduced acrossmost PPX modified surfaces (FIG. 2E).

Gene expression, measuring the level of Interleukin-6 (IL-6) cytokine ofRAW cells, further denotes capacity of PPX films to release sequesteredtherapeutic agents (FIG. 3A). Baseline results indicate all PPX surfaces(PPX-Oxd 8.6±7.3) (PPX 1.9±1.1 and PPX•2.6±0.8, P<0.01) are welltolerated and do not impose further deleterious effects when compared toControl expression (6.8±4.5). Deliberate inflammation, throughlipo-polysaccharide (LPS) stimulation (+) elevated expression levels ofIL-6, Control (+) 629.7±150.1 (P<0.001), PPX-Oxd (+) 1,126.3±301.2(P<0.05), PPX (+) 612.4±100.7 (P<0.01) and PPX•(+) 655.0±206.2 (P<0.05).DEX suppression of LPS stimulation reduces IL-6 expression when added toControls (+) DEX 217.4±54.0 (P<0.001) or integrated into PPX films(PPX•(+) DEX 209.8±34.1 (P<0.05)). Surface and chemical analysis (FIGS.3B-E & 4) completed on PPX surfaces in addition to current clinicallyavailable surgical barriers Repel™ and Preclude™ provided a basis forcomparison. The balance of hydrophilic to hydrophobic content, orsurface wettability, was obtained by measuring water contact angles(FIG. 3B-C). Native PPX surfaces displayed a mildly hydrophobic surfacewith angles of 84.08±1.33 compared to Control angles (63.58±0.62).Modified PPX surfaces PPX-Oxd and PPX-Oxd• demonstrated an increasedhydrophilic surface from plasma oxidation, 68.73±1.88 and 65.10±2.08,respectively. Clinical films Repel™ and Preclude™ showed contrastingcontent, as Repel™ revealed an extremely hydrophilic surface, 40.32±2.39compared to the very hydrophobic Preclude™ (121.77±3.64). AFMmeasurements examined surface roughness in nanometers (nm) (FIG. 3D)revealed topographical features among the various films examined. PPXsurfaces showed uniform roughness across modified and unmodifiedsurfaces, PPX-Oxd•5.76±0.51 nm and PPX 5.70±0.39 nm. Despite doublingthe amount of polymer deposited (20 gm deposition PPX-Oxd• to 40 gmdeposition PPX) no observable difference was noted in roughness values(5.76±0.51 nm to 5.70±0.39 nm). Comparatively, Repel™ and Preclude™ showcontrasting roughness values. The resorbable Repel™ film revealed aconsistent roughness of 9.36±4.35 nm; however, Preclude™ revealed aconsiderably high roughness value (63.74±40.78 nm) coupled withsubstantial variations across its surface. Film profile characterizationin the form of thickness measurements are shown in microns (μm) (FIG.3E). Modification of PPX films (PPX-Oxd and PPX-Oxd•) did not increasethe film thickness compared to base PPX depositions, (7.55 ±0.20 μm and7.76±0.07 μm to 8.38±0.09 μm). Due to the nature of plasma oxidation,the PPX surface is etched away during the process, thus resulting in theslight loss of surface material. Upon increasing the amount of PPXdeposited, 20 gm to 40 gm there is a corresponding increase in filmthickness, PPX* 17.30±0.48 μm from PPX 8.38±0.09 μm. Repel and Precludefilms revealed notably thicker profiles with values of 140.84±0.73 μmand 363.01±23.18 μm, respectively.

AFM images, shown in FIG. 6C depict surface features and compositionaluniformity among the surfaces examined. Visualization of phase images,portions A & C, in FIG. 6C, reveal surfaces composed of uniformmaterial, which was an expected result with the polymers underinvestigation. Images in B & D in FIG. 6C show topographical output as ameasure of elevation changes across the films. This information wasanalyzed and interpreted previously in FIG. 3D as RMS roughness acrosseach surface.

Chemical analysis identifying the predominant constituent elementswithin each film was measured through XPS (FIG. 4A-E). Modification ofPPX surfaces (PPX-Oxd) (FIG. 4C) show the impact of plasma oxidation inregard to the change in oxygen content from native PPX (FIG. 4B).Compared to the elemental content of control surfaces, which areconsidered biocompatible (FIG. 4A), modified PPX (PPX-Oxd) surfaces beara striking resemblance in carbon to oxygen ratios. The Repel™ film (FIG.4D) displayed an opposing ratio of carbon to oxygen content, wherebyoxygen surpassed the carbon present. Contrastingly, the Preclude™ (FIG.4E) membrane did not contain the typical oxygen carbon relationship a,but consisted mainly of fluorine with traces of carbon.

FIG. 4F-J depicts FT-IR analysis revealing chemical groups present oneach of the film surfaces. Examination of results obtained depict subtlechanges to PPX modified films (PPX-Oxd) (FIG. 14H) when compared tonative PPX (FIG. 4G) surfaces, specifically in and around the 3000 cm⁻¹peak. Plasma oxidation is a common method of prepping control surfacesfor cellular compatibility which would appear to slightly affect thecarbon-hydrogen arrangement of the aromatic backbone of either polymer.Repel™ and Preclude™ readouts correspond to their chemical compositionshown FIG. 4I-J.

In Vivo Testing

After sternotomy and adhesion treatment, the heart of 30 rabbits wastreated based on the randomized assigned group: Control group, oxidizedPPX (PPX-Oxd), PPX-Oxd infused with DEX (PPX-Oxd[DEX]), native PPX(PPX), and PPX infused with DEX and AMIO (PPX[AMIO, DEX]). Two rabbitshad a technical issue early on with displacement of the endotrachealtube during the operation which led to death. One Control rabbit wasundergoing electrophysiologic testing at the time of inadvertantendotracheal displacement and as a result only had a single (successful)arrhythmia induction attempt. Thus, 28 rabbits (93%) survived theinitial surgery with no postoperative complications. After 4 weeks,surviving rabbits (93%) were intubated, a repeat sternotomy wasperformed to observe and quantify the presence of inflammation,adhesions and measure arrhythmia induction (FIG. 5).

Gross Examination

In the Control group, highly dense adhesions formed throughout theretrosternal region between the epicardium and the sternum whichresulted in difficult dissection between the sternum and epicardium,leading to injury to the myocardium in some rabbits. In contrast,visually less dense adhesions were present in the PPX, PPX-Oxd[DEX], andPPX[AMIO, DEX] with a clear, smooth epicardium that facilitateddissection. Some rabbits appeared to lack any adhesions, similar to theappearance at the initial sternotomy. In the PPX-Oxd group, theadhesions formed were nearly similar and dense as present in the Controlgroup. Control rabbits exhibited tenacious adhesion formation resultingin an adhesion score of 2.75±0.42. PPX films, loaded with DEX exhibiteda decrease in adhesion scores compared to Controls (PPX-Oxd[DEX]0.58±0.49 and PPX[AMIO, DEX] 0.25±0.42). Adhesion scores for PPX filmsalone, modified PPX-Oxd and unmodified PPX show contrasting results dueto surface modification of each respective film (PPX-Oxd 2.83±0.41 vs.PPX 0.38±0.48, P<0.01) None of the groups had adhesions located at theposterior or lateral portions of the heart. Furthermore, the regionaldistribution of the adhesions within the heart was uniform between theapex, middle, and base of the heart.

Microscopic Analysis

All specimens had a neotissue fibrosis (NTF) layer compromised ofcollagen and adipose tissue that formed upon the myocardium (FIG. 5B-C).Tissue sections display efficacy of integrated therapeutic DEX andcellular response to PPX films (FIG. 5B-C). Control and PPX-Oxd imagesreveal significant cellular infiltration indicative of a chronicinflammatory response (FIG. 5B) in addition to the formation of aprominent NTF, 1.78±0.13 mm and 1.83±0.17 mm respectively (FIG. 5C). DEXintegrated PPX films, (PPX-Oxd [DEX], PPX [AMIO, DEX]); exhibited areduced cellular infiltration in addition to diminished NTF formation,0.93±0.09 mm and 0.80±0.10 mm (P<0.001) respectively. Conversely, nativePPX films displayed a dense cell infiltration lending to a sustainedinflammatory response (FIG. 5B); however, subsequent NTF formation wascomparable (0.88±0.10 mm, P<0.001) to DEX infused films (FIG. 5C). Thisinformation lends precedence to the ability of PPX to separatecontiguous tissue surfaces effectively, reducing adhesion formation,despite persistent inflammation.

Electrophysiology Studies

A total of 13 out of 30 rabbits underwent electrophysiologic testing.The primary arrhythmia outcome was duration of atrial fibrillation (AF)per successful induction. Rabbits with PPX[AMIO, DEX] (n=6) filmssustained atrial fibrillation for a 2.8 seconds (±1.2 sec) persuccessful induction versus 103.2 seconds (±178 sec) in Control rabbits(p=0.004)(n=7). More attempts per successful induction were required toinduce atrial fibrillation in rabbits with PPX[AMIO, DEX] films thanrabbits with Control films (13.5±7.0 vs. 5.2 ±4.3 attempts persuccessful induction, p=0.04). The duration of atrial fibrillation perrabbit was lower in PPX[AMIO, DEX] rabbits than in Control rabbits(9.5±6.8 sec vs. 187.6 ±174.7 sec of atrial fibrillation per rabbit,p=0.003). Rabbits with PPX[AMIO, DEX] films and Control rabbits did notdiffer in the total time each rabbit was exposed to induction current(180±63 vs. 125±109 seconds, p=0.253), suggesting that both populationshad equally rigorous attempts at arrhythmia induction. One rabbit with aPPX[AMIO, DEX] film was not inducible for atrial fibrillation after 32induction attempts. A PPX[AMIO, DEX] rabbit without inducible AF issupportive of the fact that the PPX[AMIO, DEX] film is protectiveagainst atrial fibrillation. If this rabbit were included, it wouldstrengthen the findings; however, because no arrhythmia was induced,this rabbit was excluded from analysis of arrhythmia.

Discussion

In-vitro assessment of PPX surfaces by time lapse images (FIGS. 6A-B),viability (FIGS. 2A, D), proliferation assays (FIGS. 2B, E) and IL-6gene expression (FIG. 3A) lend support to PPX variants as biocompatibleinterfaces. From a cellular standpoint, unmodified PPX films preventedor inhibited complete interaction of either cell line examined. Theseresults are confirmed via adhesion assays completed (FIGS. 2C, F). Theinability of macrophage or fibroblast adherence to the surfacecorrelates to a diminished capacity of in-vivo tissues from interactingor adhering to the PPX film, thereby preventing the onset of adhesionformation. This aspect lends precedence to the ability of PPX films torespond accordingly when implanted in-vivo. Consequently, those surfaceswhich facilitated adhesion may sustain the inflammatory response andresult in the formation of adhesions. Film surfaces modified to reduceor prevent cellular involvement may allow for proper regeneration ofdamaged tissues while inhibiting adhesion formation. Cytokine expression(FIG. 3A) of Interleukin-6 (IL-6) reflects the impact of therapeuticintegration in PPX films. Integration of DEX and subsequent releasereveal the suppression of intentional inflammation, throughlipo-polysaccharide (LPS) induction. Decreasing IL-6 expression acrossDEX samples exemplify the therapeutic potential of drug integration.Furthermore, basal expression of IL-6 on Control and various PPX films,indicate the compatibility of macrophages upon each surface.

Comparative chemical analysis of species present on each surfaceidentified certain chemical groups which may be conducive in preventingtissue adhesion. Data obtained (FIG. 3B-C) indicate a moderate degree ofhydrophobic content (PPX angles 84.08±1.33), appears to impede cellularor tissue interaction to the film surface. Alternatively, modificationof similar PPX surfaces when oxidized (PPX-Oxd) increased the extent ofhydrophilic groups (PPX-Oxd ange 68.73±1.88). The increase inhydrophilic content is directly attributed to an increase in oxygen onthe surface (FIG. 4C). Such surface alteration leads to profound changesin cellular growth and adhesion patterns, as noted in FIGS. 6A-B, 2C,and F. Modification through plasma oxidation increasing oxygen contentappears to alter cellular response.

Additional physical properties measured included surface roughness (FIG.3D) and film thickness (FIG. 3E). Native PPX films present smoothsurfaces and a thinner profile when compared to clinical films Repel™and Preclude™. Each PPX surface, PPX-Oxd and PPX, presented similarroughness values 5.76±0.51 nm to 5.70±0.39 nm. Additionally, changes inmaterial bulk properties may impact the inflammatory and/or healingresponse. Subsequent reduction in implanted material or bulk profilescan be considered beneficial as to lessen the disturbance to thesurrounding tissue architecture.

The PPX film infused with DEX significantly reduced the extent ofinflammation, (FIG. 5A, PPX-Oxd[DEX] 0.58±0.49 and PPX [AMIO, DEX]0.25±0.42 (P<0.001)) interpreted via adhesion formation, when comparedto Control animals 2.75±0.42. This reduction in inflammation may beattributed to DEX release from the PPX film. As noted with modified PPXsurfaces, oxidized PPX (PPX-Oxd) caused substantial adhesions(2.83±0.41). Oxidation of the surface allowed for tissue adherence,affirmed by in-vitro data, during periods of inflammation which wasinhibited through local DEX administration. Surface modification orchemical composition may exert an influence upon inflammation of tissuesurfaces as it relates to adhesion formation. Unmodified PPX surfaces,revealed a significant reduction in adhesion score 0.38±0.48 (P<0.01)without therapeutic involvement. Thus, native PPX surfaces present asurface ideal for the reduction of sustained perioperative inflammation.Tissue interaction or adherence to an implanted barrier appears todirectly lead to or perpetuate the inflammatory response resulting inthe formation of adhesions. Thus fabrication of a material surface whichinhibits tissue interaction coupled with the gradual release of ananti-inflammatory could have a profound effect at reducing chronicperioperative related inflammation.

Tissue sections obtained from each condition examined revealedinflammation trends (FIG. 5B-C) corresponding to reported adhesionscores. Infusion of DEX into PPX films, PPX-Oxd[DEX] and PPX[AMIO, DEX],resulted in minimal cellular infiltration (FIG. 5B) or inflammationcorrelating to NTF thickness (FIG. 5C) values of 0.93±0.09 mm and0.80±0.10 mm, respectively. In contrast, PPX-Oxd and Control animalsrevealed significant cellular infiltration with subsequent correspondingNTF thickness measurements of 1.78±0.13 mm and 1.83±0.17 mm. Native PPXfilms implanted without DEX appear to sustain a subdued level ofinflammation represented by cellular infiltration remaining at time ofresternotomy (FIG. 5B). However, NTF formation is reduced, 0.88±0.10 mm,(FIG. 5C) due to the film preventing interaction of opposing tissuesurfaces. These results indicate the capacity of an anti-adhesivebarrier, which does not promote or interact with tissue surfaces, toprevent chronic perioperative adhesions in lieu of therapeuticinvolvement. Historically, the most common means of preventing acuteperioperative inflammation is the transient separation of contiguoustissue surfaces with resorbable barriers. However, this methodology doesnot appear to adequately reduce chronic perioperative inflammation. Thepresence of a non-resorbable barrier which inhibits cellular interactionof opposing tissues can significantly inhibit the onset of chronicinflammation and allow for proper mesothelial response and repair at thesite of injury.

Postoperative atrial fibrillation continues to complicate surgicalintervention. Systemic amiodarone has been used as a peri-operativetherapy; however, concerns of systemic toxicity remain and releasingAMIO in a site specific manner may be a more effective method ofdelivery.31-31. Preferably, localized delivery via a DDP would reducetoxicity concerns of systemic injection. Atrial fibrillation was moredifficult to induce in rabbits with PPX[AMIO, DEX] films and moredifficult to sustain (FIG. 5D). These results indicate the ability ofPPX films to deliver a drug to the heart, thereby reducing entrance intoand duration of AF events as they might occur postoperatively.

In certain embodiments, the PPX film targets two major complications ofcardiac surgery in a combinatorial fashion through the delivery oftargeted therapeutics to the applied organ diminishing the necessity forintravenous infusion. The nano-structured film allows for the efficientrelease of therapeutics (e.g., DEX and AMIO) and remains as a protectivebarrier between the heart and chest cavity. Reduction of surgicalcomplications through application of novel nano-structured PPX filmsuccessfully suppressed peri-operative inflammation as well as atrialfibrillation. As such, a PPX loaded device is a potent tool forclinicians to mitigate these challenges. This intervention isaccomplished via twofold mechanisms of a physical barrier in addition tothe release of therapeutic agents. Additionally, it employs physicalcharacteristics which inhibit adhesion of fibrotic tissues to thesurface of the film, while also allowing for visualization of underlyingtissue.

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All publications and patents mentioned in the present application areherein incorporated by reference. Various modification and variation ofthe described methods and compositions of the invention will be apparentto those skilled in the art without departing from the scope and spiritof the invention. Although the invention has been described inconnection with specific preferred embodiments, it should be understoodthat the invention as claimed should not be unduly limited to suchspecific embodiments. Indeed, various modifications of the describedmodes for carrying out the invention that are obvious to those skilledin the relevant fields are intended to be within the scope of thefollowing claims.

We claim:
 1. An article of manufacture comprising: a polymer film with atop surface and a bottom surface, wherein said polymer film consistsessentially of a para-xylene polymer, wherein said polymer film has athickness between 1 μm and 75 μm, wherein substantially all of said topsurface has a water contact angle between 75 and 95 degrees, whereinsubstantially all of said bottom surface has a water contact anglebetween 75 and 95 degrees, and wherein said polymer film is sterilizedand sized to serve as a tissue separator and/or adhesion barrier in asubject.
 2. The article of manufacture of claim 1, wherein substantiallyall of said top surface is non-oxidized.
 3. The article of manufactureof claim 1, wherein said top surface has a carbon to oxygen ratio ofabout 2:1 to about 18:1.
 4. The article of manufacture of claim 1,wherein said top surface has a RMS roughness of about 1.0 nm to about10.0 nm.
 5. The article of manufacture of claim 1, wherein said topsurface of said polymer film has a surface area between 10 cm2 and 400cm2.
 6. The article of manufacture of claim 1, wherein para-xylenepolymer comprises parylene C.
 7. The article of manufacture of claim 1,wherein said para-xylene polymer is selected from the group consistingof: parylene A, parylene AM, parylene AF4, parylene SF, parylene HT,parylene X, parylene N, and parylene D.
 8. The article of manufacture ofclaim 1, wherein said subject is a human.
 9. The article of manufactureof claim 1, wherein substantially all of said bottom surface isnon-oxidized.
 10. The article of manufacture of claim 9, whereinsubstantially all of said bottom surface has a Carbon to Oxygen ratiorange of about 2:1 to 18:1 and RMS roughness between about 1.0 nm and10.0 nm.
 11. The article of manufacture of claim 1, wherein said polymerfilm has a thickness between 5 μm and 25 μm.
 12. A method for separatingareas in an internal region of a subject comprising: inserting anarticle of manufacture into an internal region of a subject betweenfirst and second areas, wherein said article of manufacture is asrecited in claim
 1. 13. The method of claim 12, wherein said internalregion is said subject's thoracic cavity, wherein said first area issaid subject's heart, and said second area is said subject's sternum.14. The method of claim 12, wherein said internal region is saidsubject's cephalic region, wherein said first area is said subject'soccipital area, and said second area is said subject's orbital area. 15.The method of claim 12, wherein said internal region is said subject'svertebral region, wherein said first area is said subject's cutaneoustissue, and said second area is said subject's spinal cord.
 16. Themethod of claim 12, wherein said internal region is said subject'scervical region, wherein said first area is said subject's vertebralarea, and said second area is said subject's mental area.
 17. The methodof claim 12, wherein said internal region is said subject's abdominalcavity, wherein said first area is said subject's visceral surfaces, andsaid second area is said subject's peritoneal surfaces.
 18. The methodof claim 12, wherein said internal region is said subject's pelviccavity, wherein said first area is said subject's visceral surfaces, andsaid second area is said subject's peritoneal surfaces.
 19. The methodof claim 12, wherein said internal region is said subject'sabdominopelvic cavity, wherein said first area is said subject'svisceral surfaces, and said second area is said subject's peritonealsurfaces.
 20. The method of claim 12, wherein said internal region issaid subject's abdominal cavity, wherein said first area is saidsubject's parietal peritoneum incision site or surgical wound, and saidsecond area is said subject's visceral organs or surfaces.
 21. Themethod of claim 12, wherein said internal region is said subject'spelvic cavity, wherein said first area is said subject's parietalperitoneum incision site or surgical wound, and said second area is saidsubject's visceral organs or surfaces.
 22. A system comprising: a) anarticle of manufacture as recited in claim 1, and b) a packagingcomponent, wherein said article of manufacture is located inside saidpackaging component.
 23. The system of claim 22, wherein said article ofmanufacture is sealed inside said packaging component such that saidarticle of manufacture remains sterile while inside said packagingcomponent.